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Detekce výskytu Hymenoscyphus fraxineus pomocí lapačů spor a molekulárně biologických metod v Hackerově školce
Hrnčiříková, Jana
Bachelor thesis deals with the pathogenic fungus Hymenoscyphus fraxineus, casual agent of a disease called ash dieback, which is causing major problems in stands in Czech Republic on trees growing in urban environment and in open landscape since 2004. Research in this thesis was performed in Hacker's tree nursery in mixed stand with infected ash trees. The spore trap was under an individual Fraxinus excelsior. The volumetric spore trap was used to catch the spores. Trapping took place from April 25. to November 15. 2015. Molecular biological methods were used to the subsequent quantification of the amount of spores. The research was focused on finding connections between the weather variables i.e. air temperature, air humidity and leaf moisture and the occurrence of spores during the season.
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Prostorové šíření vzdušného inokula patogenní houby Fusarium circinatum na infikované lokalitě v Galícii
Janoš, Patrik
The aerial inoculum of the Fusarium Circinatum, a fungus from the family of Ascomycota, which causes the Pine Pitch Cancer (PPC), is one of the primary disseminators of the destructive tree disease in forest stands and nurseries. Despite the fact that this pathogen is widely known nowadays, its biological characteristics are to some extent still largely unexplored, even after its introduction to Europe and the subsequent emerging of newly infected areas. In an attempt to shed more light on this topic, we conducted an experiment in the Pinus monterey and Pinus pinaster forest stands in Galicia (Spain) that were infested with the pathogen. The objective was to provide a detailed description of the spatial dissemination of the F. circinatum spores depending on the orientation and the velocity of wind. To detect and quantify the aerial inoculum of the pathogen, rotational spore traps combined with quantitative real-time PCR were employed. In three out of four conducted observations, the impact of the wind orientation with the velocity of 5 m/s manifested as a positive factor for the dissemination. The inoculum presence was detected within the distance of 1000 meters from the infected area following the draft of the wind. Our research has therefore confirmed that the wind orientation and velocity positively influence the dissemination of the pathogen spores. At the same time the study demonstrated that the combination of rotational spore traps and qPCR can be an eligible tool for the detection of F. circinatum. The results of the experiment were affected by low concentration of inoculum during the time period of the experiment, as well as by lowered reliability of the data concerning the orientation and velocity of wind.
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Methods for storing and archiving zoological tissue collections, and identification based on DNA barcoding
Aghová, Tatiana ; Benda, Petr ; Brejcha, Jindřich ; Dolejš, Petr ; Kyralová, Eva ; Mlíkovský, Jiří ; Moravec, Jiří ; Šanda, Radek ; Štundl, Jan ; Tkoč, Michal ; Vondráček, Dominik
Fulltext:  PDF; PDF; PDF; PDF
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Detekce výskytu Hymenoscyphus fraxineus pomocí lapačů spor a molekulárně biologických metod v Hackerově školce
Hrnčiříková, Jana
Bachelor thesis deals with the pathogenic fungus Hymenoscyphus fraxineus, casual agent of a disease called ash dieback, which is causing major problems in stands in Czech Republic on trees growing in urban environment and in open landscape since 2004. Research in this thesis was performed in Hacker's tree nursery in mixed stand with infected ash trees. The spore trap was under an individual Fraxinus excelsior. The volumetric spore trap was used to catch the spores. Trapping took place from April 25. to November 15. 2015. Molecular biological methods were used to the subsequent quantification of the amount of spores. The research was focused on finding connections between the weather variables i.e. air temperature, air humidity and leaf moisture and the occurrence of spores during the season.
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Detekce výskytu Hymenoscyphus fraxineus pomocí lapačů spor a molekulárně biologických metod ve Vranovickém lese
Smetanová, Daniela
This bachelor thesis deals with ash dieback. It is caused by Hymenoscyphus fraxineus pathogen. The thesis is focused on the evaluation of the connection between climatic conditions and the occurence of inoculum of this pathogenic fungi. The practical in the field consists o fair sampling. The research area was in the forest in Vranovice. The traps of Burkard´s type were used during the collection. Data were processed in the laboratory after the collection, where DNA was extracted and qPCR applied in order to quantify DNA in samples.
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Methods of Carnivores genetic monitoring in Holarctis
Smutný, Matyáš ; Černá Bolfíková, Barbora (advisor) ; Miroslav, Miroslav (referee)
Genetic monitoring begins to be widely used for monitoring carnivores, in Europe this method is also used for monitoring of other species. In this work I focused on exact definition of genetic monitoring, what is the practical use of this method and function together with other methods of monitoring. In my work I further examined invasive and non-invasive methods of gathering samples and well-known but also less known methods of monitoring of small carnivores in general and large carnivores namely.
For practical part I extracted DNA from thirty three samples of faeces and eight samples of tissue. Thirty blood samples used in this work were extracted in the research From Wolves to Dogs and Back: Genetic Composition of the Czechoslovakian Wolfdog by Smetanová et al. (2015). Scats came from Beskydy and Kokořínsko, tissues were from legal blast in Slovakia and specimen killed in Valašské Meziříčí and Kokořínsko, and blood came from dogs. After extraction I measured the concentration of DNA by spectrophotometer NanoDrop 2000 and using statistical program Statistica 12 I compared their quality. The blood had the highest average concentration od DNA, the lowest in contrast had tissue. In this work I discuss which genetic material is most suitable, in terms of yield of genetic monitoring, and additionally the limits of each method.
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DNA Extraction of Sweet Cherry (Prunus avium L.)
Hřebcová, Kateřina ; Sedlák, Petr (advisor) ; Korecký, Jiří (referee)
Isolation of high quality DNA in satisfactory yield and purity is a fundamental and essential step for all molecular-biological studies and analyses. The process of its extraction can be complicated by many of materials like are polyphenols, polysaccharides, proteins and other metabolites that can be co-isolated with nucleic acids and can act as inhibitors of PCR and cause deterioration of samples for further analyses.
In this thesis, mostly used methods of plant DNA isolation were mapped, and, in experimental part, results, regarded to the yield and purity, of selected plant DNA isolation methods were compared. DNA was obtained from various tissues of Prunus avium L. species, namely from fresh leaves, buds and from frozen embryos of several varieties. Comparison of the two commercial isolation kits (DNeasy Plant Mini Kit by Qiagen and GeneEluteTM Plant Genomic DNA Miniprep Kit, Sigma Aldrich) was the original intention. The first of the kits was replaced by simple and quick DEP-25 DNA Extraction Kit, Top-Bio and the experiment was extended with CTAB DNA isolation protocol, both with and without application of RNase into the protocol.
The results obtained proved quite significant differences between the methods used, both in yield and purity. The original assumption, supported by several studies, that commercial kits not always gain relevant results, regarded to ability to provide pure DNA, was not accurately proven, the assumption that the CTAB protocol can gain satisfactory results according to the DNA yield and purity was proved only with some tissues. The results of the spectrophotometry were supported with polymerase chain reaction (PCR) analyses conducted with the isolated DNA samples and after statistical evaluation were discussed.
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